Abstract: The milk fat globule membrane (MFGM), a trilayer membrane structure encapsulating milk fat, was embedded with various functional membrane proteins. In this study, three sample groups, MFG, MFGM, and MFGM protein, were prepared and subjected to simulated gastrointestinal digestion. Using peptidomics combined with high-throughput molecular docking, antioxidant peptides derived from MFG proteins were efficiently screened. Results showed that a total of 104, 211, and 93 MFG protein-derived peptide sequences were identified in the three groups, respectively. High-throughput molecular docking was performed with Kelch-like ECH-associating protein 1 (Keap1) as the receptor to screen peptides with potential antioxidant activity from the hydrolysates. Based on binding energy ranking, eight peptides with potential antioxidant activity were selected and synthesized via solid-phase peptide synthesis. The antioxidant activity of these peptides was evaluated using DPPH and ABTS⁺ radical scavenging assays, revealing that five peptides (PPIRPPFYPP, FPPPPPPYFPE, LRWFAGKQ, PPDWDQRP, and GYDFWYQPR) exhibited high antioxidant activity. Among them, LRWFAGKQ demonstrated the highest ABTS⁺ radical scavenging activity (IC₅₀=0.025±0.001 mg/mL), while FPPPPPPYFPE showed the highest DPPH radical scavenging activity (IC₅₀=0.374±0.010 mg/mL). This study simulated the gastrointestinal digestion of MFG proteins and efficiently screened antioxidant peptides using peptidomics and high-throughput molecular docking, laying a theoretical foundation for the potential application of MFG as a food-derived protein source for antioxidant peptide production.